1. Selection of Leafy Green Vegetable Varieties for a Pick-and-eat Diet Supplement on ISS Kennedy Space Center Team: Gioia Massa 1, Jeff Richards 2, LaShelle Spencer 2, Mary Hummerick 2, Gary Stutte 2 and Raymond Wheeler 1 Johnson Space Center Team: Grace Douglas 1 and Takiyah Sirmons 3 1 NASA, 2 Engineering Services Contract-Vencore, 3 Wyle Integ. Sci. & Eng- Lockheed Martin

2. NASA Human Research Program (HRP) Advanced Food Technology Project Safe, nutritious food that meets spaceflight requirements Sensory acceptability and variety Balance with resource constraints

3. HRP Long Duration Food System Research Plan 3 Continuing research to improve prepackaged system Supplement with pick and eat salad crops

4. Pick and Eat Crop Selection Study: Aim: Pick and Eat Crop Selection Study: Aim: To down select crop candidates for crew to grow, pick, and eat on ISS to supplement a packaged diet. ISS crop production would likely use the Veggie hardware or a similar growth system. ISS Cmdr. Swanson harvesting lettuce from the Veggie test– June, 2014 Veggie is currently on ISS in the Columbus module. ISS Cmdr. Swanson harvesting lettuce from the Veggie test– June, 2014

5. “Leafy Green” Crops Studied: ‘Tyee’ spinach ‘Flamingo’ spinach ‘Outredgeous’ Red Romaine lettuce ‘Waldmann’s Dark Green’ Leaf lettuce ‘Bull’s Blood’ beet ‘Rhubarb’ Swiss chard ‘Tokyo Bekana’ Chinese cabbage Mizuna

6. Selection Criteria Overview Horticultural factors Germination, ease of growth, amount of growth (food), size Dietary factors Percent dry matter Elemental Factors - Composition of key elements (K, Fe, Ca, Mg) Nutrient Factors - Beneficial phytonutrients (Vitamin K, Lutein, Zeaxanthin, Antioxidants) Organoleptic factors 9-pt Hedonic Scale: Overall taste, Appearance, Color, Bitterness, Flavor, Texture 5-point Just About Right Scale: Crispness, Tenderness

7. Weighting and Ranking Weighting factors were developed for each parameter based on subjective importance. Data were normalized, weighted, and ranked. Divided into: Horticultural Factors Elemental Factors Nutrient Factors Organoleptic Factors 8 varieties were down selected to 4 for nutrient and organoleptic evaluation. 4 crops were grown and shipped to JSC for organoleptic testing. Final ranking performed on down selected crops.

8. Growth Studies ConditionDayNight Photoperiod16 hours8 hours Temperature23°C18°C Test 1 23°C Test 2 Relative Humidity70% CO 2 1000 ppm Light - CWF~400 µmol·m -2 ·s -1 - Plants grown in a controlled environment chamber at KSC In test 2 eliminated thermoperiod to be similar to Veggie

12. Elemental Analysis CaFeKMg (μg/g DM) Beet14787.05 a,b 286.51 a 30416.27 a 25843.99 a Chinese cabbage17068.20 b 6.79 b 20036.45 b 7993.75 b Spinach (Flamingo) 8493.08 a,c,d 47.52 b 12854.27 b 9966.16 b Mizuna 14140.41 a,b 16.73 b 16964.88 b 6761.41 b Lettuce (Outredgeous) 8168.93 c,d 14.02 b 36169.85 a,c 6541.36 b Swiss chard 14332.48 a,b 65.02 b 26887.21 a,d 17808.77 c Spinach (Tyee) 8601.98 d 36.60 b 22427.80 a,b 10990.26 b Lettuce (Waldman’s Green) 5916.18 d 20.85 b 23656.95 a,b 4467.57 b

13. CropPlant Growth Factors Plant Growth Rank Elemental Factors Elemental Rank Plant Growth + Elemental PG+E rank 'Bull's Blood' beet 2.3083.8026.106 'Tokyo Bekana' Chinese cabbage 6.2613.3649.62 1 'Flamingo' spinach 2.5271.9484.468 Mizuna5.0522.6867.73 3 'Outredgeous' lettuce 4.2343.7337.96 2 'Rhubarb' Swiss chard 3.4553.9417.39 4 'Tyee' spinach2.9462.8355.777 'Waldmann's Green' lettuce 4.8232.3177.125

14. Nutrient Analysis ORACLuteinZeaxanthinVit. K μmol TE/gmg/g mg/100g BeetND*ND Chinese cabbage156.41 a 2.28 a 0.02 a 0.65 a Spinach (Flamingo)ND Mizuna121.84 a 3.87 b 0.02 a 2.32 b Lettuce (Outredgeous)1119.59 b 3.16 b 0.02 a 0.16 c Swiss chard1107.80 b 5.94 c 0.04 a 0.79 d Spinach (Tyee)ND Lettuce (Waldmann’s Green) ND

15. Organoleptic Factors Attribute Chinese cabbage ‘Mizuna’‘Outredgeous’ lettuce Swiss chard Overall Acceptability 7.23 (1.42)6.89 (1.18)6.69 (1.79)6.51 (2.12) Appearance8.10 (1.05)7.29 (1.73)7.66 (1.53)8.51 (0.89) Color Intensity 8.34 (0.94)8.03 (1.18)7.77 (1.46)8.57 (0.85) Bitterness6.03 (1.52)6.23 (1.46)6.14 (1.54)6.09 (1.50) Flavor7.11 (1.64)6.83 (1.36)6.51 (2.13)6.14 (2.58) Texture7.97 (1.12)7.31 (1.32)6.66 (1.97)7.31 (1.63) 9-point Hedonic Scale Attribute Chinese cabbage ‘Mizuna’‘Outredgeous’ lettuce Swiss chard Crispness3.0 (0.0)2.94 (0.23)2.46 (0.51)2.97 (0.17) Tenderness3.0 (0.0)2.94 (0.34)3.31 (0.47)2.94 (0.42) 5-point Just About Right Scale (with 3=Just About Right) Numbers are means (Standard Deviations) of N=35 tasters 1= Like Extremely, 9=Dislike Extremely

16. CropNutrientsNutrient Rank Organol. Factors Organol. Rank OverallOverall Rank 'Tokyo Bekana' Chinese cabbage 0.6348.05118.30 1 Mizuna2.1525.64215.52 3 'Outredgeous ' lettuce 1.9631.93411.85 4 'Rhubarb' Swiss chard 4.9214.55316.86 2 Final Normalized Ranking

17. Next Steps Prepare top candidates for flight testing in Veggie Grow top candidates in Veggie in plant pillows Microbiology assessment Seed sanitation Red and Blue LED light testing Work to manifest and fly Similar down selection underway with dwarf pepper and tomato varieties

18. Thank you!

19. Backup

20. ParameterWeightingRationale Plant Growth Factors Germination x 1.5Germination indicates how easy plants are to grow. SPAD x 0SPAD is a factor of plant growth (chlorophyll content) but not important for diet or a yield parameter Volume x 1.5 (inverted) Volume is important because it gives information on how much space the crop will occupy and it is a constraint for spaceflight FM x 2Fresh mass indicates crop yield - a principal factor Days to maturity x 1.5 (inverted) Indicates how quickly plants could be grown % moist x 1 (inverted)Percent moisture indicates amount of dry mass ~calories Elemental Factors Ca x 1Calcium is important but desired amount remains unclear Fe x 1.5 (inverted) Too much iron can cause issues so low iron is desired K x 2Space diet is deficient in Potassium - a principal factor Mg x 1.5More Magnesium is desirable

21. ParameterWeightingRationale Nutrient Factors ORAC x 1.5Antioxidants may help protect from radiation damage Lutein x 1.5Lutein is potentially important for eye health Zeaxanthin x 1.5Zeaxanthin is potentially important for eye health Vitamin K x 1.5The space diet is deficient in Vitamin K Organoleptic Factors Overall taste x 2Overall taste is a principal factor Appearance x 1Appearance is not important as long as it is acceptable Color x 1Color is not important as long as it is acceptable Bitter x 1Bitterness might be good or bad Flavor x 1.5Flavor is important Texture x 1Texture is not important as long as it is acceptable Crispness x 1Crispness is not important as long as it is acceptable Tenderness x 1Tenderness is not important as long as it is acceptable